Molecular tests for malaria that can be performed with unprocessed samples

可以使用未处理的样本进行疟疾分子检测

• 批准号: 8110641
• 负责人: Huimin Kong
• 金额: $ 26.96万
• 依托单位: BIOHELIX CORPORATION
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-15 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8110641
• 关键词: Address; Anti-malarial drug resistance; Antimalarials; Biological Assay; Blood; Cause of Death; Chemistry; DNA; DNA amplification; Detection; Development; Diagnostic; Diagnostic tests; Gold; Immunoassay; Laboratories; Lateral; Malaria; Microscopy; Molecular; National Institute of Allergy and Infectious Disease; Nucleic Acids; Parasites; Patients; Performance; Pharmaceutical Preparations; Phase; Plasmodium; Plasmodium falciparum; Population; Relative (related person); Resistance; Resources; Rural; Sampling; Small Business Innovation Research Grant; Staging; Technology; Testing; Whole Blood; asexual; base; fighting; helicase; inhibitor/antagonist; internal control; manufacturing process; pathogen; public health relevance

DESCRIPTION (provided by applicant): This SBIR-AT-NIAID Phase I application addresses the problem of malaria diagnostics in resource- limited settings. BioHelix, has developed an isothermal nucleic acid amplification chemistry called helicase dependent amplification (HDA) that is tolerant of the DNA amplification inhibitors present in whole blood. This proposal will focus on the application of this technology to the detection of malarial asexual parasites without nucleic acid extraction. Based on our preliminary results, the expected sensitivity of the species-specific molecular test (~100 nucleic acid copies/5L blood) better than that possible with current rapid tests for asexual stages (200 copies of parasite/5L blood). Our specific objectives for this 2-year-long project are: 1. Develop an HDA assay to detect P. falciparum, P. vivax, P. ovale, and P. malariae. 2. Develop competitive internal controls (CIC) for the species-specific assay, as well as a 2-plex assay to detect the CIC with the targets in whole blood. 3. Develop manufacturing processes for the proposed assay kit that will enable field-use of the technology. 4. Evaluate the performance of the assay relative to the gold-standard tests. 5. Obtain CE marking for the assay. 6. Submit a pre-IDE to FDA as a preliminary step to seeking FDA clearance for the test. PUBLIC HEALTH RELEVANCE: As malaria is the leading cause death in endemic regions, the current treatment approach is not based on laboratory, or microscopy testing because establishing quality-assured microscopy in rural and resource- poor settings is difficult. The liberal use of anti-malarial drugs that results from such practices presents a problem in that resistance to first line drug treatments is wide-spread. This SBIR-AT-NIAID proposal focuses on the development of a species-specific for the detection of Plasmodium in blood. More sensitive, species- specific, and stage-specific diagnostic tests will help focus drug treatment, and may slow the spread of anti- malarial drug resistance in the pathogen population. Based on our preliminary results, the expected sensitivity of the species-specific DNA test (~100 nucleic acid copies/5L blood) will be better than that possible with current rapid tests for asexual stages (200 copies of parasite/5L blood). A more sensitive test that can be performed in the field near patients would greatly enhance malaria fighting activities. We anticipate tests in the range of $7-$10 for the HDA tests should be possible, if total test volumes reach 200,000.

描述（由申请人提供）：此SBIR-AT-NIAID阶段应用程序解决了资源有限设置中疟疾诊断问题的问题。 Biohelix已开发出一种等温核酸扩增化学，称为解旋酶依赖性扩增（HDA），该化学耐受性DNA扩增抑制剂全血中存在。该建议将集中于该技术在没有核酸提取的疟疾无性寄生虫检测中的应用。基于我们的初步结果，在当前对无性阶段的快速测试（200份寄生虫/5L血液）的快速测试中，物种特异性分子检测（〜100核酸拷贝/5L血液）的预期灵敏度要好。我们对这个为期2年的项目的具体目标是：1。开发HDA测定法以检测Falciparum，P。vivax，P。Ovale和P. Malariae。 2。开发具有特定物种特异性测定的竞争性内部对照（CIC），以及2个PLEX测定法，以检测全血的靶标的CIC。 3。为拟议的测定套件开发制造过程，该工具将实现该技术的现场使用。 4。评估测定相对于金标准测试的性能。 5。获得测定的CE标记。 6.向FDA提交id，作为寻求FDA清除测试的初步步骤。 公共卫生相关性：由于疟疾是地方性地区的主要原因死亡，因此当前的治疗方法不是基于实验室或显微镜测试，因为在农村和资源不良的环境中建立质量鉴定的显微镜是困难的。由于对第一行药物治疗的抵抗力，抗疟疾药物的自由使用呈现出一个问题。该SBIR-NIAID提案重点是特异性物种特异性的发展，以检测血液中的疟原虫。更敏感，特异性和特定阶段的诊断测试将有助于聚焦药物治疗，并可能减缓抗降低耐药性在病原体种群中的传播。基于我们的初步结果，在当前对无性阶段的快速测试（200份寄生虫/5L血液）中，物种特异性DNA检测（〜100核酸拷贝/5L血液）的预期灵敏度将比结果更好。可以在患者附近的野外进行的更敏感的测试将大大增强疟疾的战斗活动。如果总测试量达到200,000，则预计HDA测试的测试范围在$ 7- $ 10的范围内。